"

From 2010.igem.org

Module I, DT and Fan's part:

Using the newly synthesized primers go through a pre-test experiment.

PCR system (FastPFU):

 H2O	        31.4μl
 5×buffer	10μl
 dNTP	        5μl
 primer1	1μl
 primer2	1μl
 template	1μl
 FastPFU	0.6μl
 Total	        50μl

Module I, Danyang's part:

Conduct PCR to amplify eGFP, Kan and Chlr with DNA polymerase pyrobest.

PCR system (FastPFU):

 H2O	        36.7μl
 10×buffer	5μl
 dNTP	        5μl
 primer1	1μl
 primer2	1μl
 template	1μl
 FastPFU	0.3μl
 Total	        50μl

result

The concentration of the M, K and C are a little lower. We’ll amplify them once again with two degrees lower of the annealing temperature.